概述

  • 产品名称Anti-JNK2抗体[EP1595Y]
    参阅全部 JNK2 一抗
  • 描述
    兔单克隆抗体[EP1595Y] to JNK2
  • 经测试应用适用于: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues near the C terminus of human JNK2.

  • 阳性对照
    • HeLa cell lysate; human breast carcinoma tissue.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

性能

应用

Our Abpromise guarantee covers the use of ab76125 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/100 - 1/500.
WB 1/1000 - 1/10000. Predicted molecular weight: 48 kDa.
IP Use a concentration of 5 µg/ml.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/40. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Responds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating a number of transcription factors, primarily components of AP-1 such as c-Jun and ATF2 and thus regulates AP-1 transcriptional activity. In T-cells, JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells.
    JNK2 isoforms display different binding patterns: alpha-1 and alpha-2 preferentially bind to c-Jun, whereas beta-1 and beta-2 bind to ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms. JUNB is not a substrate for JNK2 alpha-2, and JUND binds only weakly to it.
  • 序列相似性Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • 结构域The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • 翻译后修饰Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme. Autophosphorylated in vitro.
  • Information by UniProt
  • 数据库链接
  • 别名
    • c Jun kinase 2 antibody
    • C Jun N terminal kinase 2 antibody
    • c-Jun N-terminal kinase 2 antibody
    • JNK 55 antibody
    • JNK-55 antibody
    • JNK2 alpha antibody
    • JNK2 antibody
    • JNK2 beta antibody
    • JNK2A antibody
    • JNK2alpha antibody
    • JNK2B antibody
    • JNK2BETA antibody
    • Jun kinase antibody
    • MAP kinase 9 antibody
    • MAPK 9 antibody
    • Mapk9 antibody
    • Mitogen activated protein kinase 9 antibody
    • Mitogen-activated protein kinase 9 antibody
    • MK09_HUMAN antibody
    • P54a antibody
    • p54aSAPK antibody
    • PRKM9 antibody
    • Protein kinase, mitogen-activated, 9 antibody
    • SAPK alpha antibody
    • SAPK antibody
    • SAPK1a antibody
    • Stress activated protein kinase 1a antibody
    • Stress-activated protein kinase JNK2 antibody
    see all

Anti-JNK2 antibody [EP1595Y] 图像

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling JNK2 with purified ab76125 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).

    Control: PBS only



  • Predicted band size : 48 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: JNK2 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: MCF7 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab76125 observed at 54 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab76125 was shown to specifically react with JNK2 when JNK2 knockout samples were used. Wild-type and JNK2 knockout samples were subjected to SDS-PAGE. ab76125 and ab8245 (loading control to GAPDH) were diluted 1/2500 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • JNK2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to JNK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76125.

    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).

    Band: 48kDa; JNK2

  • ab76125 at 1/100 dilution staining JNK2 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.
  • Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/50000 dilution + HeLa cell lysate at 10 µg

    Secondary
    goat anti-rabbit-HRP at 1/1000 dilution
    Developed using the ECL technique

    Predicted band size : 48 kDa
    Observed band size : 54 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 46 kDa (possible isoform).
  • ICC/IF image of ab76125 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab76125, neat) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab76125 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76125, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-JNK2 antibody [EP1595Y] (ab76125)参考文献

This product has been referenced in:
  • Peng Y & Zhang L Activation of the TLR1/2 pathway induces the shaping of the immune response status of peripheral blood leukocytes. Exp Ther Med 7:1708-1712 (2014). WB ; Human . Read more (PubMed: 24926371) »
  • Yan T  et al. Luteolin inhibits behavioral sensitization by blocking methamphetamine-induced MAPK pathway activation in the caudate putamen in mice. PLoS One 9:e98981 (2014). WB ; Mouse . Read more (PubMed: 24901319) »

See all 6 Publications for this product

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